Antifungal bh890

ABSTRACT

A new antifungal compound, designated BH890, is produced by cultivating a new strain of Streptomyces misionensis NRRL 3609. The new antifungal can be used to inhibit growth of a variety of fungi, as, for example, Candida albicans and Cryptococcus neoformans.

United States Patent Martin et al.

ANTIFUNGAL B11890 Inventors: John Henry Edward James Martin, New City, N.Y.; John Norman Porter, Ramsey, NJ.

Assignee: American Cyanamid Company,

Stamford, Conn.

Filed: April 14, 1969 Appl. No.: 815,610

US. Cl ..424/119, 424/120 1111. C1. ..A6lk 21/00 Field 01 Search ..424/1 19, 120

[ 1 Oct. 24, 1972 Primary Examiner-Albert T. Meyers Assistant ExaminerDaren M. Stephens AttorneyNorton S. Johnson ABSTRACT A new antifungal compound, designated BH890, is produced by cultivating a new strain of streptomyces misionensis NRRL 3609. The new antifungal can be used to inhibit growth of a variety of fungi, as, for example, Candida albicans and Cryptococcus neoformans.

3 Claims, 2 Drawing Figures PATENTE-Dnm 24 m2 SHEET 1 BF 2 0 9 (.LNHQHBd) BONVLLIWSNVHL ooom oo om o3 on 08 8m 8m 89 00: 8w. 00 2 8 9 008 comm 80v 0mm OOw INVENTORS JOHN HENRY EDWARD JAMES MARTIN BY JOHN NQRMAN PORTER ATTORNEY PATENIEDOCT 24 I972 sum 2 OF 2 m. N 9 m m N w n v m w I.

03 09. com 0 88m 89 oo oo N 8100 2 008 Son 008 0mm 89 00mm ooov flzovrozmaommm IYVENTORS. JOHN HENRY EDWARD JAMES MARTIN JOHN NORMAN PORTER BY ATTURIVE) 3,700,769 1 v 2 AN' IIEUNGAL image ge) to natuial 2 dc) to Slate Ten 2 ig) to Covert Gray This invention relates to a new antifungal, to its ::l;?n l: :l? (3 lg)'spomlanonhght to heavy production by fermentation, to methods for its p g p m recovery and concentration from crude solutions, and' 5 SOLUBLE PlGMENTS to processes for its purification. The novel products are active against a variety of fungi, and the effects of the Soluble plgmems not formed on the medla used' new antifungal on specific microorganisms, together I REVERSE COLOR with its chemical and physical properties, differentiate it f previously described antifungals l ln yellowlsh to grayish to dark greenish shades.

The new antifungal, which we have designated Bl-l890, is formed during the cultivation under con- MISCELLANEOUS PHYSIOLOGICAL REACTIONS trolled conditions of a streptomycete isolated from a Nitrates reduced to mites in Organic nitrate broth; forest soil sample collected in Pennsylvania. A viable complete liquefaction of gelatin in 14 y melanoid culture of the new microorganism has been deposited l p g not produced on p p g tolerates with the Culture Collection Laboratory, Northern P 7 P Nacl growth medium Carbon Utilization Research and Development Division, Source utilization, acwl'ding t Pl'idham United States Department of Agriculture, Peoria, "1., Utilization of Carbon Compoundsby some and has been added to its permanent collection. It is tinomycetale! as an Aid for Species Determination" freely available in this repository under its Accession Bacteriol- 5611074 14 method; as follows: fair No. NRRL 3609. to good utilization of l-arabinose, d-fructose, i-inositol,

The description and identification of this new lactose, d-mannitol, d-melibiose, d-rafiinose, microorganism, maintained in the culture collection of nose, Salicini d-trehalosei y and g Lederle Laboratories, Pearl River, N .Y., was supplied utililatim'l f adOIliIOl, se and o by Dr. H. D. Tresner of these laboratories.

Observations were made of the cultural, physiologi- MICROMORPHOLOGY cal and morphological features of the culture in ac- Aerial mycelium heavy, giving rise to short, compact cordance with the methods detailed by Shirling et al., spiral chains of globose to short cylindrical spores [Methods for Characterization of Streptomyces Spe- 0.5-0.6 X 0.9-1.lp.; spore surfaces smooth, as detercies. lnternat. Journ. of Syst. Bacteriol. 16:313-340, mined by electron microscopy at 8,000X. (1966)] those recommended by Pridham ct al.,v [*A On the basis of the general characteristics observed, selclfltionrgf lljedia for Maintenangg aflllaxonomicthe culture is a member of the genus Streptomyces. The Study of Streptomyces, Antibiotics Annual (1956 grayish sporulation, spiralled chains of smooth spores 1957), pp. 947-953], for the cultivation of streptomyand lack of melanoid pigment allies the culture with a cetes. Details are recorded in Tables I-IV, and a relatively large group of Streptomyces species. Howgeneral description of the culture is given below. ever, certain characteristics such as its compact spirals Underscored descriptive colors were taken from of globose to occasionally short, cylindrical spores sug- Jacobson et al., [-Col0r Harmony Manual. 3rd gests a close relationship to species of the Streptomyces edit. Container Corp. of America, Chicago l948)]. 40 hygroscopic-like complex. When comparisons were made with members of this group, it became evident AMOUNT OF GROWTH I that the new culture differed, in that it lacked the consplcuous hygroscopic nature of spore masses normally Good on most media; llght 011 P Solution associated with S. hygroscopicus. Additionally, spores agarof the latter are typically short-cylindrical and phalangiform inappearance, Tresner et al., [Morphological AERIAL MYCELIUM AND/OR EN MASiSE SPORE Spore Types in the Streptomyces hygroscopicus-like COLOR Complex. Appl. Microbiol. 15:637-639, (1967)], Aerial mycelium whitish to grayish or tan; spore .Whrg, as1l0s epf N l {RL 39 9 w e l-emost often globose.

masses in grayish ha ranging mc ygrt Ta i? A. very Striking re lt l arssrrsitases l iasl itsr i s TABLE I Cultural Characteristics of Streptomyces misionensis NRRL 3609 Incubation 14 days; Temperature: 28 0.

Amount Soluble Medium of growth Aerial mycelium and/or spores pigment Reverse color Remarks Czapeks solution Light. Aerial mycelium whitish. Sporulation light None. Colorless agar. Asparagine dextrose Good Aerial mycelium whitish, becoming Covert Tan (2 do Slate Tan (2 lg) to unnamed Reverse mottled.

agar. ge) in sporulation zones. Sporulation moderate. (18 pn) ldk. green]. Hickey and do Aerial mycelium whitish to gray, becoming Nado Bamboo (2 go) to unnamed Reverse mottled;

Tresners agar. (2 do) in sporulation zones. Sporulation (18 pn). moderate sectorlng.

1g Yeast extract agardo Aerial mycelium gray to tan, becoming Slate Tan do Adobe Brown (3 ig) to Do.

(2 lg) in sporulation zones. Sporulation heavy. Clove Brown (3 ni) to unnamed (18 pn). Knsters oatllake. do Aerial mycelium gray to tan, becoming Slate Tan .do Lt. Mustard Tan (2 ie) to Do.

(2 ig) in sporulation zones. Sporulation moderate. unnamed (18 pn). 'lomnto paste (l0 Aerial mycelium gray to tan, becoming Slate Tan do Yellow Maple (3 ng) Reverse mottled.

onlnu-nl ngnl'. (2 lg) in sporulntion zones. Sporulation light. unnamed (18 pn). l'ntitto duxtroso .do Aorlnl mycelium white to grayish becoming Covert ..do Camel (3 ie) to unnamed Reverse mottled;

:tgiu'. (tray (2 lo) in sporulution zones. Sporulution (18 pn). starch hydrolysis luoderato. light. Bennett's agar. .do Aerial mycelium gray to tan, becoming Slate Tan do Covert Brown (2 li) to Reverse mottled;

(2 ig) iusporultttion zones. Sporulation moderate. miammed (18 pn). moderate sectoring. Inorganic snlts- .(lO Aerial mycelium whitish, becoming Beige Brown do Biscuit (2 cc) to Slate Tau l)o.

starch agar. (3 lg) in sporulzttion zones. Sporulation heavy. (2 ig) to unnamed (18 pn).

species Streptomyces misionensis, Cercos et al., [Misionina: antibi6tico polinico producido por Streptomycesrmisionensis, n. sp. Revista de Investigaciones Agricolas 16:5-28 (1962)]. Good agreement was 'observed in spore color, sporophore morphology, spore shape, spore surface configuration, melanoid pigment production and carbon source utilization patterns. Furthermore, such cultural features as growth habit, color of vegetative mycelium and the mottled appearance of the culture reverses on certain media, were remarkably similar. When comparisons were made with the published descriptions and/or reference cultures of other species, none was found that more closely resembled this culture that S. misionensis. Therefore,

NRRL 3609 will, hereafter, be considered a strain of 1 Miscellaneous Physiological Reaction of Streptomyces misionensil NRRL 3609 Temperature: ZBC.

Medium Incubation Amount Physiological Reaction Period of Growth Organic 7 days Good Slight reduction Nitrate of nitrate to Broth nitrate. 7 Organic 14 days Good Nitrate reduced to Nitrate nitrate. Broth Gelatin 7 days Good Slight liquefaction Gelatin l4 days Good Complete liquefaction Peptone 24 hrs. Good Melanoid pigments lron Agar not fonned 443% NaCl 10 days Tolerated up to 7% in Yeast NaCl in growth medium Extract Agar TABLE IV Carbon Source Utilization Pattern of Streptomyces mtlrionensis NRRL 3609 Incubation: 10 days Temperature: 28C.

Carbon Source Utilization Adonital l-Arabinose d-Fructose i-lnositol Lactose d-M annitol d-Melezitose d-Melibiose uouwuwwo' d-Raffinose l-Rhamnose Saliein Sucrose d-Trehalose d-Xylose Dextrose Negative Control Ounwouwu some Utilization l-Poor Utilization Z-Fair Utilization O-No Utiliza- It is to be understood that for the production of the new antifungal the present invention is not limited to this particular organism or to organism fully answering the above growth and microscopic characteristics which are given for illustrative purposes. In fact, it is 5 desired and intended to include the use of mutants produced from the described organism by' various means, such as x-radiation, ultraviolet radiation, nitrogen mustard, phage exposure, and the like.

THE FERMENTATION PROCESS Cultivation of the organism S. misionensis may-be carried out in a wide variety of liquid culture media. Media which are useful for the production of the novel antibiotic include an assimilable source of carbon such as starch, sugar, molasses, glycerol, etc.; an assimilable source of nitrogen such as protein, protein hydrolysate, polypeptides, amino acids, corn steep liquor, etc.; and inorganic anions and cations, such as potassium, sodium, calcium, sulfate, phosphate, chloride, etc. Trace elements such as boron, molybdenum, copper, etc.; are supplied as impurities of other constituents of the media. Aeration in tanks and bottles is provided'by forcing sterile air through or onto the surface of the fermenting medium. Further agitation in tanks is provided by a mechanical impeller. An antifoarning agent, such as 1 percent octadecanol in lard oil may be added as needed.

lNOCULUM PREPARATION Shaker flask inoculum of S. mirionensis is prepared by inoculating 100 milliliters of sterile liquid medium in 500 milliliter flasks with scrapings or washings of spores from an agar slant of the culture. The following medium is ordinarily used:

Cerelose 20 grams Soy flour l0 grams Corn Steep liquor 5 grams Calcium carbonate 3 grams Water to 1,000 milliliters.

The flasks are incubated at a temperature from 25 to 29 C., preferably 28 C., and agitated vigorously on a rotary shaker for 30 to 48 hours. These 100 milliliter inocula are used to inoculate 1 liter and 12 liter batches of the same medium in 2-liter and 20-liter glass fermentors. The inoculum mash is aerated with sterile air while growth is continued for 30 to 48 hours. These batches of inocula are used to inoculate tank fermen- TANK FERMENTATION For the production of BH890 in tank fermentors the @Be t ss atl msiwi i599W!st? l5.0 grams Calcium carbonate 7 9.5 grams Ammonium sulfate 6.75 grams Casein 3.0 grams Cottonseed flour 2.5 grams Ammonium chloride 2.0 grams Manganese sulfate 0.10 grams Water to 1,000 milliliters.

Lard oil is added to the medium in the amount of 0.8 percent v/v. Each tank is inoculated with about 3 percent of the inoculum prepared as described above. Aeration is supplied at the rate of 0.5 to 1.0 liter of sterile air per liter of broth per minute and the fermenting mixture is agitated by an impeller driven at 300 to 800 r.p.m. The temperature is maintained at 2529 C., usually at 28 C. The fermentation is ordinarily continued for from 60 to 90 hours at which time the mash is harvested.

ISOLATION PROCEDURE After the fermentation is completed, the fermented mash containing the antifungal of this invention is filtered, preferably with the addition of diatomaceous earth or any other conventional filter aid. Normally the mycelial filter cake pad is washed with a small portion of water. Both filtrate and wash are discarded. The antifungal is extracted from the mycelial cake with methanol using about 1 liter of methanol for every 2.5 to 3.0 liters of liquid mash. Ordinarily, a second extraction of the cake with a smaller portion of methanol ensures complete extraction. The extracts are combined and concentrated to an aqueous phase under reduced pressure. The aqueous phase is allowed to stand overnight at 4 C. A crude solid product containing components which are referred to as BH890a and BH890B is formed on standing. The solid is collected and washed with acetone. The washed solid is first air dried and then dried over phosphorous pentoxide or other drying agent under reduced pressure.

The a component is prepared from a portion of the crude material thus prepared by repeated recrystallizations. Essentially, the B component is removed as an impurity leaving the purified a component in isolated form. The recrystallizations are carried out by dissolving a portion of crude BH890 in dimethylformamide and filtering the solution. Other solvents, as for example, dimethylacetamide and dimethylsulfoxide, also may be used. Ethyl acetate or ethyl acetate and water is used to precipitate the antifungal. Ordinarily, placing the mixture in a coldroom (temperature around 0-5 C.) tends to ensure complete precipitation. The crystalline solid is collected by filtration and washed with water and with acetone and air dried. The recrystallization procedure ordinarily is twice repeated in order to result in a purified Bl-l890a free of the B component.

The B component is prepared from a portion of crude BH890 by separating the material into the a and ,8 components by solvent countercurrent distribution.

A useful solvent system comprises ethyl acetate, secbutanol, methanol and buffer in the ratio of 900 to 303 to 160 to 600 by volume respectively. The buffer is prepared by combining 6.3 ml. of triethylamine with 2 liters of water, adjusting to pH 7.5 with glacial acetic acid and diluting to 2,400 ml. with water. With a 200- I tube counter-current distribution apparatus about 300 transfers is sufficient for good separation of the components. The progress of the separation maybe noni;

6 tored by optical density readings at 303 p. and plotting the readings against tube numbers. Appropriate fractions are combined, the liquid concentrated and lyophilized to recover both the a and the B components.

PHYSICAL CHARACTERISTICS The two components BH890a and Bh890B may be distinguished by selected physical characteristics. The analytical samples of both components are essentially ash free. BH890a was dried under reduced pressure in an Abderhalden drying apparatus over boiling acetone for 16 hours and BH890B was dried under reduced pressure over P 0 over boiling acetone overnight.

Components BH890a and BH890/3 contain the elements carbomhydrogen, nitrogen and oxygen in substantially the following percentages by weights:

8118901: 81-18905 Carbon 58.52 57.78 Hydrogen 8.73 7.74 Nitrogen 1.78 1.59 Oxygen (diff) 30.97 32.89

BH890a gives a neutralization equivalent of 936 when titrated in glacial acetic acid with perchloric acid and a neutralization equivalent of 962 when titrated in pyridine using tetrabutyl ammonium hydroxide.

The specific rotation of BH890a is determined in a variety of solvents and the values obtained are given in Table V immediately below.

TABLE V Specific Rotation of BH890a in Various Solvents Solvent .Cpncentratim 2 Dimethylformamide 0.988% 19.2" Dimethylsulfoxide 1.031% 5.8 Glacial acetic acid 0.863% 10.4 Pyridine 0.930% 18.2

The ,8 component has no definitive melting point. An infrared absorption spectrum of BH890B in a KBr pellet is prepared in the standard manner. It exhibits characteristic absorption in the infrared region of the spectrum at the following wavelengths expressed in microns: 3.00, 3.45, 5.85, 6.36, 6.70, 6.90, 7.25, 8.50, 9.40, 10.00, 11.85, 13.33, and 14.40. The infrared curve is shown in FIG. 2 of the accompanying drawings.

The B component in methanol shows ultraviolet absorption maxima at:

variety of solvents and the values obtained are given in Table VI, following.

TABLE VI Specific Rotation of BH890B in Various Solvents Solvent Concentration M1,,

Dimethylfonnamide 0.747% 4.03 Dimethylsulfoxide 0.951% 18.7 Glacial acetic acid 0.993% 8.07 Pyridine 1.038% 9.7

Antifungal BH890 is clearly distinguished from other antifungals by the characterization data given above and by its antifungal activity. The in vitro activity of this new antifungal is presented in the Table below which shows the minimal inhibitory concentration required to inhibit the growth of representative microorganisms in a nutrient medium.

TABLE VII In Vitro Antifungal Activity of BH890* Organism Minimal inhibitory Concentration mcgJml.

Candida albicans 300 E8 3 2 Cryplococcur neoforman:

SP E 1 3 8 2 Epiderm ophylon floccosum ATCC 10227 (E139) l Micrasparum audauini ATCC 14057 (15140) 2 Microrparum cam:

ATCC 10214 (E55) 5 Microsporum gypseum ATCC 14683 (E130) 2.5 Phinlophthora jeanselmei NIH 8724 (E16) 7 5 Tricllopllylan rubrum (E97) 2.5 Trichophylan lansuranr NIH 662 (E10) 2 Trichophylon mentagrophyles (Ell) 2.5

Standard agar dilution procedure Antifungal BH890 is active in vivo against Candida albicans and C ryptococcus neoformans. The new antifungal is thereby potentially useful as a therapeutic agent in treating fungal infections in mammals caused by said microorganisms. The new antifungal can be expected to be usefully employed for treating or combating such infections by topical application or parenteral a u fl watiqup whsitta Own)- The usefulness of the new antifungal is demonstrated by its ability to control systemic lethal infections by or with. ths9L 1i$minm The test with C. albicans was run with Carworth Farms CF 1 female mice, weight about 20 grams, infected intravenously with a lethal dose of a 1:20 dilution of a 24 hour trypticase soy broth culture of the organism. Theinfected mice were treated with Bh890 in varying dosages by subcutaneous injections within 1 hour after infection or subcutaneous injections within 1 hour after infection followed by a second dose 4 hours later. Record was made of the size of each group and the number of mice remaining alive 6 days after infection.

TABLE VIII Alive/Total mice, 6 days after infection Total dosage mgJmouse Single Dose Two Dose:

Infected, non-treated control mice: 0.60 Non-infected, non-treated control mice: 15/15 TABLE IX Alive/Total mice, 7 days after infection Total dosage mgJmouse Single Dose Two Doses Infected, non-treated control mice: 1/60. Non-infected, non-treated control mice: 15/15.

The invention will be described in greater detail in conjunction with the following specific examples.

TabIe T/III b eIoW illustrates antifungal activity of EXAMPLE 1 lnoculum Preparation A typical medium used to grow the primary inoculum was prepared according to the following formula:

Cerelosc 20 grams Soy flour 10 grams Corn steep liquor grams Calcium carbonate 3 grams Water to 1,000 milliliters.

Washed or scraped spores from an agar slant of S. misionensis are used to inoculate 2 500 milliliter flasks containing 100 milliliters each of the above sterile medium. The flasks are then placed on a rotary shaker and agitated vigorously for 48 hours at 28 C. The resulting flask inoculum is transferred to a 5 gallon glassj merlt r Q2t@rt. .2 me i9 the sterile medium. The glass fermentor is aerated with sterile air while growth is carried out fgr aboutitl hours, after which time the contents are used to seed a 300-liter tank fermentor.

EXAMPLE 2 Fermentation A fermentation medium is prepared according to the following formula:

Starch 52.5 grams Corn flour 14.5 grams Corn steep liquor 15.0 grams Lard oil is used in the medium in the amount of 0.8 percent v/v. The fermentation medium is sterilized at 120 C. with steam at 20 pounds pressure for 45-60 minutes. The pH of the medium after sterilization is about 6.6. Three hundred liters of sterile medium in a 400-liter tank fermentor is inoculated with 12 liters of inoculum, prepared as described in Example 1. The fermentation is carried out at 28 C. using Hodag LG-8 oil as a defoaming agent. Aeration is supplied at the rate of 0.5 liter of sterile air per liter of mash per minute. The mash is agitated by an impeller driven at about 300 revolutions per minute. At the end of approximately 66 hours of fermentation time the mash is harvested.

EXAMPLE 3 Isolation to an aqueous phase (volume about 6 liter) and this concentrate is allowed to stand overnight at 4f" C. A

birefringent solid which forms on standing is collected by centrifugation and washed with acetone. The residual acetone is removed by evaporation into the air and the solid is dried over P 0 under reduced pressure. The dried crystalline solid of crude BH890 weighs 118 grams.

EXAMPLE 4 Preparation of BH890a A portion (40 grams) of the crude BH890, as obtained from the previous example, is dissolved in ml. of-dimethylformamide and the mixture is filtered. Ethyl acetate is added to the filtrate until the cloud point is reached. About 150 ml. of water is added after which an additional 150 ml. of ethyl acetate is added with stirring. The resultant mixture is allowed to stand overnight at 4 C. The light yellow colored solid which forms and is suspended in the two phase system is collected by filtration and washed first with water and then with acetone. The air-dried solid weighs 29 grams. The crude material is recrystallized twice again by first dissolving in dimethylformarnide and repeating the above procedure from that point on. About 4.4 grams of purified BH890a having the physical and chemical characteristics hereinbefore described is ob- EXAMPLE 5 Preparation of BH890/3 Crude BH890 as obtained from Example 3, is used as starting material and separated into a and B components by solvent countercurrent distribution. The solvent system is composed of ethyl acetate, sec-but'anol, methanol and buffer (90013032160z600 by volume) wherein the buffer is prepared by stirring together triethylamine (6.3 ml.) and water (2 liters) and adjusting the pH of the solution to 7.5 with glacial acetic acid and diluting to 2,400 ml. with water. The countercurrent distribution is carried out in a 200 tube apparatus using 10 ml. of lower phase and 10 ml. of upper phase per tube and 310 transfers. The sample charge is placed in the first 15 tubes by preparing a saturated solution of BH890 in 150 ml. of lower phase and 150 ml. of upper phase, giving a net charge of 350 mg. At the completion of 310 transfers a one-half ml. portion of lower phase is withdrawn from selected tubes. Each sample is diluted with 10 ml. of methanol and its optical density at 303 p. is determined. A curve having first a minor and then a major peak is obtained by plotting optical density against tube number. Tube What is claimed is: l. A substance antibiotic BH890a, a compound which a. is effective in inhibiting the growth of fungi, and in its essentially pure crystalline form; b. has the following elemental analysis: C, 58.52; H, 8.73; N, 1.78; O, 30.97;

c. has ultraviolet rmxima at: 230 u. (E 370), 7

278p.(E 320), 289 p. (E m 616), 302 M i c1|| 318 i can d. has a characteristic infrared absorption spectrum as shown in FIG. 2. v 3. A compound selected from the group consisting of antibiotic BH89OB as characterized in claim 1 and antibiotic BH890B as characterized in claim 4.

UNHTEE ETA'IES r1: rs strict CEii'llilCzi'lE Ci CCER'EQIECN Patent No. 3,7 a-7 9 g; October 24, "1972 Inventor) John Henry Edward James Martin and John Norman Porter It is certified that error appears in the above-identified patent and that said Letters Patent are hereby corrected as shown below:

Column 1, line 51, after "(l966)]" insert Media used in the study were selected from Column 5, Table III, under the heading Physiological Reaction", the first entry should read Slight reduction of nitrate to nitrite. --5 the second entry should read Nitrate reduced to nitrite. Y

' 0/ Column 7, line 10, ;1(E 370.)" should read a Y 0 250 1(E 570) line 12, "27882(E 320)" should read 278;1(E i% 520) ---5 Table VI, under the heading "[oflg the last entry should read -9.7

Column 12, Claim 5 should read as follows: A compound selected from the group consisting of antibiotic BH890d as characterized in Claim 1 and antibiotic BH8906 as characterized in Claim 2.

Signed and sealed this 29th day of May 1973 v (SEAL) Attest:

EDWARD M.FLETCHER,JR. ROBERT GOTTSCHALK Attesting Officer v Commissioner of Patents 'ORM PO-105O (10-69) USCOMM-DC 60376-P69 i 0.5. GOVIIIIIINT PRINTING OFFICE 19G! 0-366-334 

2. A substance antibiotic BH890 Beta , a compound which a. is effective in inhibiting the growth of fungi, and in its essentially pure crystalline form; b. has the following elemental analysis: C, 57.78; H, 7.74; N, 1.59; O, 32.89; c. has ultraviolet maxima at: 230 Mu (E1%1 cm 370), 278 Mu (E1%1 cm 320), 289 Mu (E1%1 cm 616), 302 Mu (E1%1 cm 934), 318 Mu (E1%1 cm 840); and d. has a characteristic infrared absorption spectrum as shown in FIG.
 2. 3. A compound selected from the group consisting of antibiotic BH890 Beta as characterized in claim 1 and antibiotic BH890 Beta as characterized in claim
 4. 